Abstract

Alcaligenes faecalis is a heterotrophic nitrifying bacterium that oxidizes ammonia and generates nitrite and nitrate. When A. faecalis was cultivated in a medium containing pyruvate and ammonia as the sole carbon and nitrogen sources, respectively, high concentrations of nitrite accumulated in the medium whose carbon/nitrogen (C/N) ratio was lower than 10 during the exponential growth phase, while the accumulation was not observed in the medium whose C/N ratio was higher than 15. Comparative transcriptome analysis was performed using nitrifying and non-nitrifying cells of A. faecalis cultivated in media whose C/N ratios were 5 and 20, respectively, to evaluate the fluctuations of gene expression during induction of heterotrophic nitrification. Expression levels of genes involved in primary metabolism did not change significantly in the cells at the exponential growth phase under both conditions. We observed a significant increase in the expression levels of four gene clusters: pod cluster containing the gene encoding pyruvic oxime dioxygenase (POD), podh cluster containing the gene encoding a POD homolog (PODh), suf cluster involved in an iron-sulfur cluster biogenesis, and dnf cluster involved in a novel hydroxylamine oxidation pathway in the nitrifying cells. Our results provide valuable insight into the biochemical mechanism of heterotrophic nitrification.

Highlights

  • Alcaligenes faecalis is a heterotrophic nitrifying bacterium that oxidizes ammonia and generates nitrite and nitrate

  • Alcaligenes faecalis was cultivated in a synthetic medium containing 10 mM sodium pyruvate and 8 mM ammonium chloride as sole carbon and nitrogen sources, respectively, and the concentration of nitrite in the medium and the pyruvic oxime dioxygenase (POD) activity of the bacterial cells was measured

  • POD activity in the bacterial cells reached the maximum in the late exponential growth phase, rapidly decreased in the subsequent stationary phase

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Summary

Introduction

Alcaligenes faecalis is a heterotrophic nitrifying bacterium that oxidizes ammonia and generates nitrite and nitrate. Comparative transcriptome analysis using the A. faecalis cells cultivated under the low and high C/N ratio conditions, where the nitrifying activity is induced or not induced, respectively, suggested the involvement of four gene clusters in the heterotrophic nitrification process.

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